Human TNF-alpha ELISA Kit (Colorimetric)

ELISA: TNF-alpha ELISA Kit [NBP1-91170] - Standard curve

• Reactivity: Hu
• Applications: ELISA
• Suitable Sample Type: Cell culture supernatant, serum, plasma (EDTA, citrate, heparin), urine
• Standard Curve Range: 7.8 - 500 pg/mL
• Sensitivity: 2.3 pg/mL

Human TNF-alpha ELISA Kit (Colorimetric) Summary

• Description: The human TNF-alpha ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of human TNF-alpha
• Specificity: The interference of circulating factors of the immune system was evaluated by spiking these proteins at physiologically relevant concentrations into a human TNF alpha positive serum. There was no interference detected, namely not with TNF-R (60 kDa and 80 kDa).
• Standard Curve Range: 7.8 - 500 pg/mL
• Sensitivity: 2.3 pg/mL
• Assay Type: Sandwich ELISA
• Inter-Assay: CV% < 7.4%
• Intra-Assay: CV% < 6.0%
• Spike Recovery: The recovery ranged from 72% to 112% with an overall mean recovery of 93%. Please see protocol for more specific Spike Recovery details.
• Sample Volume: 50 uL
• Kit Type: ELISA Kit (Colorimetric)
• Gene: TNF

Applications/Dilutions

• Dilutions:
  • ELISA

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Alternate Names for Human TNF-alpha ELISA Kit (Colorimetric)

• APC1 protein
• Cachectin
• Cachetin
• DIF
• TNF
• TNF, monocyte-derived
• TNFA
• TNF-A
• TNFalpha
• TNF-alpha
• TNF-alphacachectin
• TNFATNF, macrophage-derived
• TNFG1F
• TNFSF1A
• TNFSF2
• TNFSF2TNF superfamily, member 2
• tumor necrosis factor (TNF superfamily, member 2)
• tumor necrosis factor alpha
• Tumor necrosis factor ligand superfamily member 2
• tumor necrosis factor
• tumor necrosis factor-alpha

Background

Tumor necrosis factor (TNF)-alpha is a pro-inflammatory cytokine belonging to the TNF superfamily that is secreted by monocytes/macrophages, T cell, and natural killer (NK), among others (1). TNF-alpha is synthesized as a 233 amino acid (aa) transmembrane protein (mTNF-alpha) with a theoretical molecular weight (MW) of 26 kDa (1,2) that forms a homotrimer. mTNF-alpha is cleaved by TNF-alpha converting enzyme (TACE) and released in its 157 aa, 17 kDa soluble form (sTNF-alpha) (1-5). Both mTNF-alpha and sTNF-alpha are capable of binding type 1 TNF receptors (TNFR1), whereas mTNF-alpha predominately binds to TNFR2 (1,2). TNF-alpha binding to its receptors causes receptor recruitment of adaptor proteins, formation of signaling complexes, and downstream signaling cascades (e.g. MAPK, NF-kappaB, and Caspase-8), leading to distinct cellular responses such as survival, proliferation, inflammation, necroptosis, and apoptosis (1-5).

TNF-alpha is critical for normal immune response; however, dysregulation of TNF-alpha production can result in various pathologies (2,4,5). Excessive production of pro-inflammatory cytokines including interleukin 1 (IL-1), IL-6, and TNF-alpha has been implicated in an array of autoimmune diseases like rheumatoid arthritis (RA), inflammatory bowel disease (IBD), and psoriasis (2,4,5). Anti-TNF monoclonal antibodies, including Infliximab, and soluble TNFR have been approved for the treatment of autoimmune and TNF-mediated diseases (5). Additionally, data suggests that TNF inhibitors can be beneficial for treating patients experiencing immune-related adverse events associated with immune checkpoint inhibitor cancer treatment (6).

References

1. Holbrook J, Lara-Reyna S, Jarosz-Griffiths H, McDermott M. Tumour necrosis factor signalling in health and disease. F1000Res. 2019;8:F1000 Faculty Rev-111. https://doi.org/10.12688/f1000research.17023.1

2. Jang DI, Lee AH, Shin HY, et al. The Role of Tumor Necrosis Factor Alpha (TNF-alpha) in Autoimmune Disease and Current TNF-alpha Inhibitors in Therapeutics. Int J Mol Sci. 2021;22(5):2719. https://doi.org/10.3390/ijms22052719

3. Horiuchi T, Mitoma H, Harashima S, Tsukamoto H, Shimoda T. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford). 2010;49(7):1215-1228. https://doi.org/10.1093/rheumatology/keq031

4. Webster JD, Vucic D. The Balance of TNF Mediated Pathways Regulates Inflammatory Cell Death Signaling in Healthy and Diseased Tissues. Front Cell Dev Biol. 2020;8:365. https://doi.org/10.3389/fcell.2020.00365

5. Kalliolias GD, Ivashkiv LB. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol. 2016; 12(1):49-62. https://doi.org/10.1038/nrrheum.2015.169

6. Chen AY, Wolchok JD, Bass AR. TNF in the era of immune checkpoint inhibitors: friend or foe?. Nat Rev Rheumatol. 2021;17(4):213-223. doi:10.1038/s41584-021-00584-4

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

Publications for TNF-alpha ELISA Kit (NBP1-91170)(5)

Publications using NBP1-91170

• Zhang Q, Bai X, Wang R et al. 4-octyl Itaconate inhibits lipopolysaccharide (LPS)-induced osteoarthritis via activating Nrf2 signalling pathway Journal of cellular and molecular medicine 2022-01-23 [PMID: 35068055]
• Yang R, Yan Y, Wu Z Et al. Resveratrol-loaded titania nanotube coatings promote osteogenesis and inhibit inflammation through reducing the reactive oxygen species production via regulation of NF-kappa B signaling pathway Materials science & engineering. C, Materials for biological applications 2021-12-01 [PMID: 34857292] (ELISA)
• Chen YJ, Lin TL, Cai Z et al. Assessment of acute pancreatitis severity via determination of serum levels of hsa-miR-126-5p and IL-6 Experimental and Therapeutic Medicine 2020-11-09 [PMID: 33262812] (ELISA, Mouse)
• Liu L, Guo H, Song A et al. Progranulin inhibits LPS-induced macrophage M1 polarization via NF-kappaB and MAPK pathways BMC Immunol. 2020-06-05 [PMID: 32503416]
• Ji L, Hou X, Liu W et al. Paeoniflorin inhibits activation of the IRAK1-NF-kB signaling pathway in peritoneal macrophages from lupus-prone MRL/lpr mice Microb. Pathog. 2018-08-24 [PMID: 30149133] (ELISA, Mouse)

FAQs for TNF-alpha ELISA Kit (NBP1-91170). (Showing 1 - 5 of 5 FAQs).

1. I need to know about TNF-alpha antibody kits for mice.
   • We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.
2. I would like to ask you for help. I need an antibody for Elisa to detect human TNF alpha in a supernatant from a cell culture (meaning supernatant after centrifugation of collected cell suspension from a plate well). Which of your antibodies against human tnf alpha would be suitable? I would like to buy only the primary antibody.
   • I would recommend catalog number NBP1-67821 or NB600-587; however, a full list of our anti-human TNF alpha antibodies suitable for use in ELISA can be found using this link.
3. I am interested in a TNF alpha antibody, cross reactive for human, rat and mouse (host species: rabbit). Could your product NBP1-19532 be used in western blotting applications? Or do you have a similar product in your catalog which could fit with my request?
   • The antibody you mention, NBP1-19532, has not yet been validated in Western blot. I would instead recommend either NBP1-67821 or NB600-587. These are both rabbit polyclonal antibodies that cross-reacts with human, mouse and rat and have been used in Western blotting.
4. Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
   • I am very sorry, but all of our products are for scientific research use only, and none are intended or approved for use in humans.
5. wondering what the difference is between your quantikine and duo set elisas?
   • Usually the duosets do not have the entire kit such as plates and buffers, whereas the other kits are complete.

Bioinformatics

• Gene Symbol: TNF